Comparative intra-articular gene transfer of seven adeno-associated virus serotypes reveals that AAV2 mediates the most efficient transduction to mouse arthritic chondrocytes

Abstract

Osteoarthritis (OA) is the most common arthropathy, characterized by progressive degeneration of the articular cartilage. Currently, there are no disease-modifying approaches for OA treatment. Adeno-associated virus (AAV)-mediated gene therapy has recently become a potential treatment for OA due to its exceptional characteristics; however, the tropism and transduction efficiency of different AAV serotypes to articular joints and the safety profile of AAV applications are still unknown. The present study aims to screen an ideal AAV serotype to efficiently transfer genes to arthritic cartilage. AAV vectors of different serotypes expressing eGFP protein were injected into the knee joint cavities of mice, with all joint tissues collected 30 days after AAV injection. The transduction efficiency of AAVs was quantified by assessing the fluorescent intensities of eGFP in the cartilage of knee joints. Structural and morphological changes were analyzed by toluidine blue staining. Changes to ECM metabolism and pyroptosis of chondrocytes were determined by immunohistochemical staining. Fluorescence analysis of eGFP showed that eGFP was expressed in the cartilage of knee joints injected with each AAV vector. Quantification of eGFP intensity indicated that AAV2, 7 and 8 had the highest transduction efficiencies. Both toluidine blue staining and Mankin score showed that AAV6 aggravated cartilage degeneration. The analysis of key molecules in ECM metabolism suggested that AAV5 and 7 significantly reduced collagen type II, while AAV9 increased ADAMTS-4 but decreased MMP-19. In addition, transduction with AAV2, 5, 7 and 8 had no obvious effect on pyroptosis of chondrocytes. Comprehensive score analysis also showed that AAV2 had the highest score in intra-articular gene transfer. Collectively, our findings point to AAV2 as the best AAV serotype candidate for gene transfer on arthritic cartilage, resulting in minimal impact to ECM metabolism and pyroptosis of chondrocytes.

α‐Solanine attenuates chondrocyte pyroptosis to improve osteoarthritis via suppressing NF‐κB pathway

“…Extensive works have highlighted the role of IL‐1β in the development of OA and the involvement of nod‐like receptor protein‐3 (NLRP3)‐mediated inflammasome in promoting various joint compartments has garnered significant attention. Both in vitro and in vivo OA model experiments conducted by our team and other researchers have consistently demonstrated that aberrant activation of NF‐κB signalling leads to crucial activation of caspase‐1‐mediated pyroptosis, which drives chondrocyte pyroptosis in OA pathogenesis 10–14 . Moreover, subchondral bone and articular cartilage act together not only mechanically but biologically to accelerate OA progression 15 .…”

“…Both in vitro and in vivo OA model experiments conducted by our team and other researchers have consistently demonstrated that aberrant activation of NF‐κB signalling leads to crucial activation of caspase‐1‐mediated pyroptosis, which drives chondrocyte pyroptosis in OA pathogenesis. 10 , 11 , 12 , 13 , 14 Moreover, subchondral bone and articular cartilage act together not only mechanically but biologically to accelerate OA progression. 15 Emerging evidence reveals that subchondral bone lesions, including angiogenesis and sensory neuron ingrowth, are responsible for cartilage sabotage and pain.…”

Cellular and Tissue Selectivity of AAV Serotypes for Gene Delivery to Chondrocytes and Cartilage

“…In this study, we showed that AAV2 and AAV6.2 could effectively deliver therapeutic genes into human joint tissues. Others have reported that AAV2 has the highest transduction efficiency in the cartilage of knee joints 19 , it has been used in clinical trials in patients with inflammatory arthritis. 20 Collectively, these studies support the current data that AAV2 is one of the most efficient vector systems with which to transfer therapeutic genes into chondrocytes of human joint cartilage.…”

AAV vectors: The Rubik’s cube of human gene therapy

“… Data compiled from (Refs. 3 , 8 , 14 , 15 , 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 ). Superscript letters: h, human; c, canine; I, pig; m, murine; p, non-human primate; r, rat; s, sheep.…”

“… Data compiled from (Refs. 3 , 8 , 14 , 15 , 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 ). …”